Lack of beta 1 integrin gene in embryonic stem cells affects morphology, adhesion, and migration but not integration into the inner cell mass of blastocysts.
Lack of beta 1 integrin gene in embryonic stem cells affects morphology, adhesion, and migration but not integration into the inner cell mass of blastocysts.
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DOI：
10.1083/jcb.128.5.979
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发表时间：
1995-03
影响因子：
7.8
通讯作者：
ALBRECHT, R
中科院分区：
文献类型：
--
作者：
FASSLER, R;PFAFF, M;MURPHY, J;NOEGEL, AA;JOHANSSON, S;TIMPL, R;ALBRECHT, R
设计了一种基因捕获型靶向载体，将β 1整合素基因在胚胎干细胞中进行了定位。使用该载体，超过50%的ES细胞克隆获得了β 1整联蛋白基因的破坏，并且单个克隆在两个等位基因中都发生了突变。纯合突变体不产生β 1整合素mRNA或蛋白质，而α 3，α 5和α 6整合素亚基被转录，但在细胞表面上检测不到。杂合突变体表现出降低的β 1表达和α/β 1异二聚体的表面定位。在任何突变体上，α V亚基表达均未受损。纯合子ES细胞突变体缺乏层粘连蛋白和纤连蛋白，但不是玻连蛋白，并显示出减少与成纤维细胞饲养层的关联。此外，它们不向成纤维细胞培养基中的化学引诱物迁移。这些功能在杂合突变体中均未受损。扫描电子显微镜显示，纯合子细胞显示较少的细胞间连接，并有许多微绒毛通常不发现野生型和杂合子细胞。这种细胞形状的深刻变化与细胞骨架成分的表达和分布的总体变化无关。出乎意料的是，显微注射到囊胚中证明了纯合和杂合突变体完全整合到内细胞团中。这将允许在几种体内情况下研究β 1整联蛋白缺乏的后果。
A gene trap-type targeting vector was designed to inactivate the beta 1 integrin gene in embryonic stem (ES) cells. Using this vector more than 50% of the ES cell clones acquired a disruption in the beta 1 integrin gene and a single clone was mutated in both alleles. The homozygous mutant did not produce beta 1 integrin mRNA or protein, while alpha 3, alpha 5, and alpha 6 integrin subunits were transcribed but not detectable on the cell surface. Heterozygous mutants showed reduced beta 1 expression and surface localization of alpha/beta 1 heterodimers. The alpha V subunit expression was not impaired on any of the mutants. Homozygous ES cell mutants lacked adhesiveness for laminin and fibronectin but not for vitronectin and showed a reduced association with a fibroblast feeder layer. Furthermore, they did not migrate towards chemoattractants in fibroblast medium. None of these functions were impaired in heterozygous mutants. Scanning electron microscopy revealed that homozygous cells showed fewer cell-cell junctions and had many microvilli not usually found on wild type and heterozygous cells. This profound change in cell shape is not associated with gross alterations in the expression and distribution of cytoskeletal components. Unexpectedly, microinjection into blastocysts demonstrated full integration of homozygous and heterozygous mutants into the inner cell mass. This will allow studies of the consequences of beta 1 integrin deficiency in several in vivo situations.
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影响因子：
14.9
作者：
LAIRD, PW;ZIJDERVELD, A;BERNS, A
通讯作者：
BERNS, A
影响因子：
64.5
作者：
MENKO, AS;BOETTIGER, D
通讯作者：
BOETTIGER, D
影响因子：
3.3
作者：
DELWEL, GO;DEMELKER, AA;SONNENBERG, A
通讯作者：
SONNENBERG, A
影响因子：
15.3
作者：
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通讯作者：
BROWN, EJ
影响因子：
5.3
作者：
JAT, PS;CEPKO, CL;SHARP, PA
通讯作者：
SHARP, PA